Crystallization and preliminary x-ray analysis of the cAMP-dependent protein kinase catalytic subunit from Saccharomyces cerevisiae

Biochemistry ◽  
1991 ◽  
Vol 30 (43) ◽  
pp. 10595-10600 ◽  
Author(s):  
Jeff Kuret ◽  
James W. Pflugrath
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protein Kinase ◽  
Catalytic Subunit ◽  
Dependent Protein Kinase ◽  
X Ray ◽  
Camp Dependent Protein Kinase ◽  
Dependent Protein

Crystallization and preliminary X-ray analysis of the unliganded recombinant catalytic subunit of cAMP-dependent protein kinase

1998 ◽  
Vol 54 (6) ◽  
pp. 1401-1404 ◽  
Author(s):  
Narendra Narayana ◽  
Pearl Akamine ◽  
Nguyen-Huu Xuong ◽  
Susan S. Taylor
Keyword(s):  
Protein Kinase ◽  
Unit Cell ◽  
Catalytic Subunit ◽  
Monoclinic Space Group ◽  
Dependent Protein Kinase ◽  
Data Set ◽  
X Ray ◽  
Cell Parameters ◽  
Camp Dependent Protein Kinase ◽  
Dependent Protein

X-ray diffraction-quality crystals of the unliganded mouse recombinant catalytic subunit of cAMP-dependent protein kinase were grown by the hanging-drop vapour-diffusion technique using 2-methyl-2,4-pentanediol as precipitant. The crystals belong to the monoclinic space group P21 with unit-cell parameters a = 48.9, b = 147.4, c = 54.2 Å, β = 110.2°. A data set to 3.0 Å resolution with 92% completeness has been collected using synchrotron radiation. The unit cell contains four molecules of molecular weight 40 kDa with a corresponding volume solvent content of 45%.

sck1, a high copy number suppressor of defects in the cAMP-dependent protein kinase pathway in fission yeast, encodes a protein homologous to the Saccharomyces cerevisiae SCH9 kinase.

Genetics ◽  
1995 ◽  
Vol 140 (2) ◽  
pp. 457-467 ◽  
Author(s):  
M Jin ◽  
M Fujita ◽  
B M Culley ◽  
E Apolinario ◽  
M Yamamoto ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protein Kinase ◽  
Adenylate Cyclase ◽  
Stationary Phase ◽  
Copy Number ◽  
Catalytic Subunit ◽  
High Copy Number ◽  
Dependent Protein Kinase ◽  
Camp Dependent Protein Kinase ◽  
Dependent Protein

Abstract Schizosaccharomyces pombe regulates intracellular cAMP levels, and thus cAMP-dependent protein kinase (PKA) activity, in response to changes in nutrient conditions. Mutations in any of eight git genes inhibit glucose repression of fbp1 transcription, alter the cell morphology, and cause a reduction in the growth rate. The eight git genes encode components of an adenylate cyclase activation pathway, adenylate cyclase itself, and the catalytic subunit of PKA. Three of these genes have been identified in other studies as regulators of meiosis. Here we show that the sck1 gene, cloned as a high copy number suppressor of a mutation in git3, is able to suppress the defects conferred by a mutation in any of these git genes. Sequence analysis suggests that sck1 encodes a protein most closely related to the Saccharomyces cerevisiae SCH9 protein kinase that had previously been identified as a high copy number suppressor of mutations in S. cerevisiae that reduce or eliminate PKA activity. Disruption of the sck1 gene causes a significant delay in exit from stationary phase when combined with a disruption of the pka1 (git6) gene encoding the catalytic subunit of PKA. However, the sck1 disruption by itself has little or no effect upon fbp1 transcription, meiosis, or exit from stationary phase, and does not enhance the constitutive fbp1 transcription observed in a pka1 mutant. Therefore, sck1 appears to function in a redundant fashion to pka1, but to varying degrees, in the pathways regulated by pka1.

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